Composition comprising at least two dry powders obtained by spray drying to increase the stability of the formulation

ABSTRACT

The present invention relates to inhalation formulations of drugs in the form of dry powder for inhalation administration deliverable as such with an inhaler and provided with high deliverability, respirability and stability. In particular, the invention relates to a pharmaceutical composition for inhalation in the form of powder comprising at least a first and a second powder, in which at least said first powder contains an active agent in an amount greater than 1% by weight with respect to the weight of said first powder. Both the powders comprise leucine in an amount ranging from 5 to 70% by weight with respect to the weight of each powder and a sugar in an amount ranging from 20 to 90% by weight with respect to the weight of each powder. The composition has a fine particle fraction (FPF) greater than 60% and a delivered fraction (DF) greater than 80.

The present invention relates to inhalation formulations of drugs in theform of dry powder for inhalation administration as such with an inhalerand having high deliverability, respirability and stability.

Inhalation therapy with aerosol preparations is used to administeractive agents to the respiratory tract, in the mucosal, tracheal andbronchial regions. The term aerosol describes a preparation consistingof particles or fine droplets carried by a gas (usually air) to the siteof therapeutic action. When the site of therapeutic action involves thepulmonary alveoli and small bronchi, the drug must be dispersed in theform of droplets or particles with an aerodynamic diameter of less than5.0 μm.

When the target is the pharyngeal region, larger particles are moreappropriate.

Conditions suitable for these treatments are represented bybronchospasm, inflammation, mucosal edema, pulmonary infections and thelike.

-   -   Currently, administration of drugs in the deep lung region is        obtained through inhalation devices such as:    -   nebulizers, in which the drug is dissolved or dispersed in the        form of suspension and carried to the lung as nebulized        droplets;    -   powder inhalers, capable of delivering the drug present in the        inhaler as dry micronized particles; or    -   pressurized inhalers, through which the drug—again in the form        of droplets of solution or suspension—is carried to the deep        lung region by an inert gas expanded rapidly in air by a        pressurized canister.

In all these cases, technological problems have been encountered in thedevelopment of effective products that still limit the administration ofdrugs by inhalation.

From a clinical point of view, an ideal inhalation product should allowdifferent administration methods to be used by the patient, since theinhalers described are generally suitable for different types ofpatients and administration conditions. In general, nebulizer therapy isprevalently used by elderly or pediatric patients, while therapy withdry powder or pressurized inhalers is more suitable for adults. However,the use of nebulizers is currently still considered effective, since thepatient inhales the drug under rest conditions and without using forcedinhalation, which is instead required for an inhalation powder. Instead,in the case of a pressurized inhaler, the product must be takencoordinating inspiration with activation of the device, to prevent thedelivered particles from impacting on the bottom of the throat andfailing to reach the deep lung.

For these reasons, the inhalation formulations used in these three typesof inhalation devices are generally essentially very different from oneanother.

In the case of products for nebulizers, formulations are substantiallyconstituted by solutions or suspensions containing as excipients salts,surfactants and preservatives to ensure isotonicity of the preparation,homogeneity of the particle size distribution in case of suspensions,and protection against microbial contamination.

In the case of pressurized preparations, the composition usuallycontains surfactants, propellants and co-solvents. In inhalationformulations in powder form, the excipients essentially consist oflactose with different particle size, used as carrier.

Some formulation or stability constraints in some cases have limitedindustrial development of inhalation products and, apart fromcorticosteroids, which exist substantially in all inhalation forms, insome bronchodilator and anti-cholinergic active agents some forms ofadministration are not available on the market. These limitations areparticularly important since current respiratory therapy makes use ofcombinations of drugs of different kinds as the most effective techniqueand, in this regard, it has been possible to develop only a small numberof corticosteroid-bronchodilator combinations, prevalently in the formof inhalation powder.

With regard to nebulized forms, the patient is left to extemporaneouslycombine different formulated products, which might even be incompatiblewith one another.

From a therapeutic point of view, it is therefore limiting for a patientnot to be able to take the same drug in different conditions, such as athome, at work, while travelling and in an emergency. In the differentsituations indicated, a patient might be obliged to use differentpreparations containing different active agents.

The most important of the formulation problems encountered in thedevelopment of inhalation products concerns chemical stability inrelation to atmospheric agents, which cause rapid degradation of theinhalation preparation and, consequently, decrease the shelf life of theproduct containing this preparation.

The stability of an inhalation product is particularly important, sinceit must be administered to the deep lung while maintaining its physicalfeatures for quantitative penetration of particles or droplets to thedeepest regions thereof. Added to this is the fact that the number ofexcipients currently approved for inhalation administration andtherefore acceptable in terms of toxicity for the pulmonary tissue isvery limited.

The literature reports examples of dry inhalation powders with highdispersibility in air due to their low density. These powders areusually formulated with a high content of phospholipids, in particulardipalmitoylphosphatidylcholine (DPPC).

A powder of this kind is described in the patent applicationUS2005/0074498 A1, relating to low density particles, with an internallyhollow morphology, obtained by spray drying with the use of surfactantsconstituted by phospholipids in combination with a blowing agent). Thehollow structure is described as resulting from the precise combinationof the blowing agent and of the surfactant phospholipid. The documentdoes not describe examples of similar morphology obtained withoutphospholipids. The use of phospholipids as surfactants determines theprincipal features of the product obtained and above all its sensitivityand stability in relation to atmospheric agents, which would beparticularly influenced in this case by moisture. Moreover, the patentliterature (US 2001/0036481 A1) indicates values of the phospholipidtransition temperature (Tg) with humidity of 41° C. for DPPC, 55° C. fordistearoylphosphatidylcholine (DSPC) and 63° C. fordipalmitoylphosphatidylethanolamine (DPPE), the three phospholipids mostcompatible with pulmonary administration.

The transition temperature is defined as the temperature required tocause a change in the physical state of the lipids, from the ordered gelphase in which the hydrocarbon chains are lying flat and closely packed,to the disordered liquid-crystalline phase in which the hydrocarbonchains are randomly oriented and fluid.

These Tg values are all much lower than the characteristic Tg value ofamorphous lactose.

It is known that the closer the Tg is to the temperature of theenvironment in which the preparation is stored, the easier thetransition will be. It is also known that in a system in which the mainexcipient is fluid and loosely packed, the molecular mobility of thecomponents is very high, and consequently has a propensity to causedifferent chemical reactions and degradation of the active agents.

Therefore, the solution of producing porous particles for inhalationadministration with phospholipids does not appear to be supported byreasonable scientific evaluation in relation to the long term stabilityof the product.

The aforesaid patent application, besides application as inhalationpowder, also describes application of these particles in an inhalerdevice with a propellant gas. This administration would be impossiblewith a conventional nebulizer by dispersing the particles in water oraqueous solution, given the incompatibility of the materials with water,above all due to their tendency to float on the surface of the liquid orto dissolve slowly therein.

The concept of “high porosity” or “low density” has been used in asubstantially equivalent manner in the cited patent applications.

In particular, the term “density” has been used not to refer to theabsolute density of the particles, since this, measured with a heliumpycnometer, would identify the density of the solid materials formingthe powder and the particles according to the equation:

ρ=P/V (g/cc)

but rather to refer to the apparent density (in some documents by othersdescribed as “envelope density”) of the particle, considering itsoverall volume.

Given the technical difficulty of measuring this overall volume for eachsingle particle, the cited patent applications have referred to volume(and subsequently to density) parameters of the powder as bulk volumeand tapped volume.

The patent application WO 03/0350030 A1 describes the preparation of akit for inhalation administration that considers the preparation of asolid dry form containing a drug prepared by freeze drying a solution.The process, also described through examples, present great difficultiesin relation to industrial production and, above all, provide noguarantees of substantial improvement of the stability of the activeagent over time. In fact, after freeze drying the drug added to theformulation is dispersed in an excipient network characterized by highporosity that cannot be modulated or modified through the process.Although it is useful from the point of view of rapid dissolution of thesolid form, this porosity increases exposure of the drug to atmosphericagents and compromises its stability. In the specific case, no data areprovided on the porosity of the freeze-dried products obtained in theexamples, but literature data obtained through indirect measurementsplace the apparent density (corresponding to the bulk density of apowder) of formulated freeze-dried tablets containing sugars andsurfactants between 0.05 and 0.2 g/cc.

The patent application CA2536319 describes a pharmaceutical compositionobtained by spray drying, with a moisture content below 1%. According towhat is indicated, this very low moisture content is necessary to ensurethe stability of the composition, as a water content of over 1% in thepowder would cause degradation of the pharmacologically activesubstances, resulting in a loss of efficacy of the composition. Toreduce the level of moisture the composition is constituted by a largeamount of mannitol, which however compromises the physical features ofthe powder considerably, increasing the particle size and decreasing thedose of powder delivered from the mouthpiece of the inhalation deviceused.

The problem of producing inhalation powders with high dispersibility hasbeen solved through the engineering of particles that contain the drugas dispersed as possible.

Briefly, the technique used is that of producing essentially fineparticles (geometric mean diameter greater than 4.0 μm) constituted bysmall amounts of active agent dispersed at molecular level inside anappropriate matrix of excipients capable of guaranteeing, through thespray drying preparation technique, the formation of a low densitycoarse particle.

This formulation approach requires the use of high percentages ofexcipients in the formulation, but enables small amounts of active agentto be contained in the composition.

For this reason, although these compositions solve the problem ofaerodynamic performance, they fail to solve significant questions interms of chemical stability.

The production of an inhalation powder in which the content % of activeagent is high using a spray drying technique must instead be consideredadvantageous in terms of chemical stability. Considering the commonactive agents of respiratory therapy, in the majority of cases thiscontent % of active agent would be too high to allow the production ofan inhalation powder form, given the limited amount of powder thatconstitutes an individual dose of product.

In fact, this amount of powder is too small to be dosed reproducibly byany industrial device for producing individual doses of inhalationpowders.

Therefore, the production of an inhalation powder that is stable bothfrom a chemical and physical point of view must necessarily reconcilethe need for stability of the active agents used with the need to ensureadequate aerosol performance in terms of deposition in the deep lung.

From the point of view of chemical stability, an ideal approach isrepresented by the production of dry powders containing large amounts ofactive agent in combination with a sugar capable of decreasing molecularmobility in the particles of powder and a hydrophobic excipient capableof limiting interaction with the external environment and absorption ofwater by the powder.

From the point of view of aerosol performance, the same powder must becharacterized by an adequate particle diameter for inhalationadministration and by a composition capable of facilitating particledisaggregation at the time of inhalation.

At the same time, convergence of physical composition features of thepowder must coincide with the ability to divide the powder evenly usingdevices for the industrial preparation of products in the form ofinhalation powder in individual doses or of multidose inhalers capableof drawing a relatively large dose from a storage chamber containedtherein.

In the light of all of the aforesaid considerations, it would beadvantageous to be able to produce a pharmaceutical composition forinhalation use in the form of dry powder that is stable and easy toadminister with common dispensers for inhalation powders, whileremaining easy to produce.

It would also be advantageous to obtain a solid composition in the formof dry powder, which can be used as diluent of inhalation powders inorder to enable correct mixing of powders containing different activeagents also in small amounts and at the same time maintains highstability of the formulation, preventing degradation of the activeagents.

However, the problem of providing an inhalation formulation of drugsthat is stable and administrable with common dispensers of inhalationpowders, with features of high deliverability and respirability, andwhich can be produced with a commercially viable process, currentlyremains unsolved or unsatisfactorily solved.

According to the present invention, formulation is a combination of twoor more different powders obtained according to the preparationprocedure described by mixing, and HLSA and HLDA powders are powder witha high loading of active which are made according to the preparativespray drying procedure.

In the present description the wording “pharmaceutical composition” and“formulation” have the same meaning.

Therefore, a first aspect of the present invention is to provide apharmaceutical composition for inhalation characterized in that itcomprises at least a first and a second powder, in which at least saidfirst powder comprises an active agent in an amount greater than 1% byweight with respect to the weight of said first powder, said first andsecond powder containing:

-   -   a) leucine in an amount ranging from 5 to 70% by weight with        respect to the weight of each powder;    -   b) a sugar in an amount ranging from 20 to 90% by weight with        respect to the weight of each powder;        in which said composition has a fine particle fraction (FPF)        greater than 60% and a percentage of the dose delivered from the        mouthpiece (DF) greater than 80%.

Another aspect of the invention is represented by a process forpreparing said solid pharmaceutical composition comprising the followingsteps:

-   -   a) providing at least a first powder obtained by spray drying        comprising an active agent in an amount greater than 1% by        weight with respect to the weight of the powder, leucine in an        amount ranging from 5 to 70% by weight with respect to the        weight of the powder, a sugar substantially amorphous after        obtaining the powder by spray drying in an amount ranging from        20 to 90% by weight with respect to the weight of the powder;    -   b) providing a second powder obtained by spray drying comprising        leucine in an amount ranging from 5 to 70% by weight with        respect to the weight of the powder, a sugar in an amount        ranging from 20 to 90% by weight with respect to the weight of        the powder;    -   c) mixing the powders.

A further aspect of the invention is represented by a Kit foradministration of a drug as inhalation powder, comprising a dosed amountof the composition according to the present invention and an inhalationdevice.

Another aspect of the present invention is represented by a solidcomposition for use as diluent of inhalation powders comprising apowder, characterized in that it comprises:

-   -   a) leucine in an amount ranging from 5 to 70% by weight with        respect to the weight of the powder;    -   b) a sugar in an amount ranging from 20 to 90% by weight with        respect to the weight of the powder;        in which said composition has an aerosol fine particle fraction        (aerodynamic diameter <5.0 μm), greater than 60% and a        percentage of the dose delivered from the mouthpiece (DF)        greater than 80%.

According to the present invention, the term “active agent” is intendedas any substance with a desired biological therapeutic efficacy.

Examples of active agents that can be administered by inhalationcomprise: β₂ agonists; steroids such as glucocorticosteroids (preferablyanti-inflammatory agents); anti-cholinergic agents; leukotrieneantagonists; inhibitors of leukotriene synthesis; mucolytics;antibiotics, pain relievers in general such as analgesic andanti-inflammatory agents (including steroid and non-steroidanti-inflammatory agents); cardiovascular agents such as glucosides;respiratory agents; anti-asthma agents; bronchodilators; anti-canceragents; alkaloids (i.e. rye ergot alkaloids) or triptans such assumatriptan or rizatriptan that can be used to treat migraine; agents(i.e. sulfonylurea) used to treat diabetes and related dysfunctions;sleep inducing drugs such as sedative and hypnotic agents; psychicenergizers; appetite inhibitors; anti-arthritis agents; anti-malariaagents; anti-epileptic agents; anti-thrombotic agents; anti-hypertensiveagents; anti-arrhythmic agents; anti-oxidant agents; anti-psychoticagents; anxyolitics; anti-convulsant agents; anti-emetic agents;anti-infective agents; anti-hystamines; anti-fungus and anti-viralagents; drugs to treat neurological dysfunctions such as Parkinson'sdisease (dopamine antagonists); drugs to treat alcoholism and otherforms of addiction; drugs such as vasodilators to treat erectiledysfunction; muscle relaxants; muscle contractors; opioids; stimulatingagents; tranquilizers; antibiotics such as macrolides; aminoglycosides;fluoroquinolones and β-lactames; vaccines; cytokines; growth factors;hormones including birth-control drugs; sympathomimetic agents;diuretics; lipid regulating agents; anti-androgen agents;anti-parasitics; blood thinners; neoplastic agents; anti-neoplasticagents; hypoglycemic agents; nutritional agents and supplements; growthsupplements; anti-enteric agents; vaccines; antibodies; diagnostic andcontrast agents; or mixtures of the above substances (e.g. combinationsfor the treatment of asthma containing steroids and β-agonists).

The aforesaid active agents belong to one or more structural classes,including, but not limited to, small molecules (preferably smallinsoluble molecules), peptides, polypeptides, proteins, polysaccharides,steroids, nucleotides, oligonucleotides, polynucleotides, fats,electrolytes and the like.

Specific examples include β₂ agonists salbutamol, salmeterol (e.g.salmeterol xinafoate), formoterol and formoterol fumarate, fenoterol,steroids such as beclomethasone dipropionate, budesonide, fluticasone(e.g. fluticasone propionate). In relation to peptides and proteins, thepresent invention also includes synthetic, recombinant, native,glycosylated and non glycosylated peptides and proteins, biologicallyactive fragments and analogs.

Active agents for which an immediate release into the bloodstream isparticularly advantageous to obtain a rapid pharmacological effectinclude those to be used to treat migraine, nausea, insomnia, allergicreactions (including anaphylactic reactions), neurological andpsychiatric disorders (in particular panic attacks and other psychosesor neuroses), erectile dysfunction, diabetes and related diseases, heartdiseases, anti-convulsive agents, bronchodilators and active agents totreat pain and inflammation. According to the present invention,vaccines constituted by antibodies, cells, corpuscles and cellularportions can also be administered.

Other examples of active substances are steroids and their salts, suchas budesonide, testosterone, progesterone, flunisolide, triamcinolone,beclomethasone, betamethasone, dexamethasone, fluticasone,methylprednisolone, prednisone, hydrocortisone and the like; peptidessuch as cyclosporine and other water-insoluble peptides; retinoids suchas cis-retinoic acid, 13-trans-retinoic acid and other derivatives ofvitamin A and of beta-carotene; vitamins D, E and K and their precursorsand water-insoluble derivatives; prostaglandins, leukotriens and theiractivators and inhibitors including prostacyclin, prostaglandins E1 andE2, tetrahydrocannabinol, pulmonary surfactant lipids; lipid-solubleanti-oxidants; hydrophobic antibiotics and chemotherapic drugs such asamphotericin B, adriamycin and the like.

In particular, according to the present invention the active agent is adegradable active agent, i.e. a substance capable of undergoingdegradation processes as a function of the amount of water present inthe formulation.

According to the present invention, the term “sugar” is intended asmonosaccharides with 5 or more carbon atoms, disaccharides,oligosaccharides or polysaccharides and also polyols with 5 or morecarbon atoms (often also defined as sugar-alcohol)

Examples of sugars that can be administered by inhalation comprise:lactose, threalose, sucrose, maltose, melibiose, cellobiose, mannitol,dextrins, maltodextrins, sorbitol, galactitol, iditol, volemitol,fucitol, inositol, maltitol, lactitol, isomalt, maltotriitol,maltotetraitol, polyglycitol. The amount of sugar present in the powderscontained in the pharmaceutical composition of the present descriptionranges from 20 to 90% by weight with respect to the weight of eachpowder, is preferably present in an amount ranging from 20 to 80% byweight with respect to the weight of each powder, even more preferablyin an amount ranging from 40 to 80% by weight with respect to the weightof each powder.

According to the present invention the powders contained in thepharmaceutical composition of the present description include ahydrophobic substance to reduce moisture sensitivity. This hydrophobicsubstance is leucine, which also facilitates particle disaggregation.Leucine is present in an amount ranging from 5 to 70% by weight withrespect to the weight of each powder. Preferably the amount of leucinepresent in the powders contained in the pharmaceutical compositionranges from 15 to 70% by weight with respect to the weight of eachpowder, even more preferably from 18 to 55% by weight with respect tothe weight of each powder.

According to the present invention the first and the second powder thatconstitute the composition comprise a surfactant in an amount rangingfrom 0.2 to 2% by weight with respect to the weight of each powder,preferably in an amount ranging from 0.4 to 0.8% by weight with respectto the weight of each powder.

The surfactant of the pharmaceutical composition according to theinvention can be selected from the various classes of surfactants forpharmaceutical use.

Surfactants suitable to be used in the present invention are all thosesubstances characterized by medium or low molecular weight containing ahydrophobic moiety, generally readily soluble in an organic solvent butweakly soluble or totally insoluble in water, and a hydrophilic (orpolar) moiety, weakly soluble or completely insoluble in an organicsolvent but readily soluble in water. Surfactants are classifiedaccording to their polar moiety. Therefore, surfactants with anegatively charged polar moiety are called anionic surfactants, whilecationic surfactants contain a positively charged polar moiety.Uncharged surfactants are generally called non ionic, while surfactantswith both a positive and negative charge are called zwitterionic.Examples of anionic surfactants are represented by the salts of fattyacids (better known as soaps), sulphates, sulphate ethers and phosphateesters. Cationic surfactants are frequently based on polar groupscontaining amino groups. The most common non ionic surfactants are basedon polar groups containing oligo-(ethylene-oxide) groups. Zwitterionicsurfactants are generally characterized by a polar group constituted bya quaternary amine and a sulphuric or carboxylic group.

Specific examples of this application are represented by the followingsurfactants: benzalkonium chloride, cetrimide, docusate sodium, glycerylmonooleate, sorbitan esters, sodium lauryl sulphate, polysorbates,phospholipids, biliary salts.

Non ionic surfactants, such as polysorbates and polyoxyethylene andpolyoxypropylene block copolymers, known as “Poloxamers”, are preferred.Polysorbates are described in the CTFA International Cosmetic IngredientDictionary as mixtures of sorbitol and sorbitol anhydride fatty acidesters condensed with ethylene oxide. Particularly preferred are nonionic surfactants of the series known as “Tween”, in particular thesurfactant known as “Tween 80”, a polyoxyethylene sorbitan monooleateavailable on the market.

The presence of a surfactant, and preferably of Tween 80, is necessaryto reduce the electrostatic charges found in powders without it, flow ofthe powder and maintenance of a homogeneous solid state without initialcrystallization.

According to the present invention, the term “inhalable” is intended asa powder suitable for pulmonary administration. An inhalable powder canbe dispersed and inhaled by means of an appropriate inhaler, so that theparticle can enter the lungs and alveoli to provide the pharmacologicalfeatures of the active agent of which it is formed. A particle withaerodynamic diameter of less than 5.0 μm is normally consideredinhalable.

The term “amorphous” according to the present invention is intended as apowder that contains less than 70% of crystalline fraction, morepreferably less than 55%. The pharmaceutical composition described inthis text has a ratio between the amount of powder in amorphous formthat constitutes the composition expressed by weight and the amount ofsugar present in the composition expressed by weight ranging from 0.8 to2.0. This ratio indicates that the sugar present in the powder is asubstantially amorphous sugar, which therefore has a crystallinefraction of less than 50%. This enables the sugar to coordinate thewater present in the composition, preventing it from being available tohydrolyze the active agent, thereby making it ineffective.

In a further embodiment, the pharmaceutical composition according to thepresent invention comprises a third powder comprising an active agent,according to the previously indicated features, in an amount greaterthan 1% by weight with respect to the weight of said third powder,leucine in an amount ranging from 5 to 70% by weight with respect to theweight of said third powder, a sugar in an amount ranging from 20 to 90%by weight with respect to the weight of said third powder.

With a pharmaceutical composition as described in this secondembodiment, it is possible to obtain a pharmacologically activecomposition that can comprise the combination of two or more differentactive agents capable of acting synergically, or simply actingsimultaneously in the application site, so as to reduce the number ofadministrations.

The term “fine particle fraction (FPF)” is intended as the fraction ofpowder, with respect to the total delivered by an inhaler, which has anaerodynamic diameter (dae) of less than 5.0 μm. The characterizationtest that is performed to evaluate this property of the powder is theMulti Stage Liquid Impinger (MSLI) test, as described in the EuropeanPharmacopoeia current ed. The conditions for performing this testconsist in subjecting the powder to an inhalation through the inhalersuch as to generate a flow of 60 litres/min. This flow is obtained byproducing a pressure drop of 2 KPa in the system.

The term “delivered fraction (DF)” is intended as the fraction of activeagent, with respect to the total loaded, delivered by a powder inhalerin standard inhalation conditions.

The characterization test performed to evaluate this property of thepowder is the DUSA test, as described in the European Pharmacopoeiacurrent ed. The conditions for performing this test consist insubjecting the powder to an inhalation through the inhaler such as toproduce a pressure drop of 4 KPa in the system.

The preferred production process of the powder according to theinvention is spray drying starting from a solution of leucine, of asugar and a surfactant in which the drug, if present, is dissolved ordispersed as suspension or emulsion.

The preferred particle size for this powder provides that at least 50%of the size distribution (X50) is below 5 μm, preferably below 3 μm,more preferably below 2.0 μm, also to increase the surface areaoptimizing deep lung deposition.

According to the present invention, the powder that constitutes thepharmaceutical composition is a substantially dry powder, i.e. a powderwith a moisture content of less than 10%, preferably less than 5%, morepreferably below 3%. This dry powder preferably has no water capable ofhydrolyzing the active agent making it inactive. The amount of moisturepresent in the composition is controlled by the presence of leucine,which limits the content due to its hydrophic features, both in the stepto produce the powder and in the subsequent handling steps, and ofsugar, which traps the water in a structure that becomes increasinglyrigid over time, preventing the water from hydrolyzing the active agent.

The process for preparing the pharmaceutical composition according tothe invention substantially comprises the operations of:

-   a) providing at least a first powder obtained by spray drying    comprising an active agent in an amount greater than 1% by weight    with respect to the weight of the powder, leucine in an amount    ranging from 5 to 70% by weight with respect to the weight of the    powder, a sugar substantially amorphous after obtaining the powder    by spray drying in an amount ranging from 20 to 90% by weight with    respect to the weight of the powder;-   b) providing a second powder obtained by spray draying comprising    leucine in an amount ranging from 5 to 70% by weight with respect to    the weight of the powder, a sugar substantially amorphous after    obtaining the powder by spray drying in an amount ranging from 20 to    90% by weight with respect to the weight of the powder;-   c) mixing the powders.

In particular the production process of the composition, in step a) andb) of obtaining the powders by spray drying, consists of a series ofoperations illustrated below: For step a):

-   -   preparing a first phase (A) in which an active agent is present        in an appropriate liquid medium;    -   preparing a second phase (B) in which the leucine, the sugar and        surfactants are dissolved or dispersed in an aqueous medium;    -   mixing said phases (A) and (B) to obtain a third phase (C) in        which the liquid medium is homogeneous;    -   drying said phase (C) in controlled conditions to obtain a dry        powder with particles having a size distribution with median        diameter of less than 10.0 μm;    -   collecting said dry powder.

Phase (A) can be either a suspension of the active agent in an aqueousor non aqueous medium, or a solution of the active agent in anappropriate solvent.

Preparation of a solution is preferable, and the organic solvent isselected from those soluble in water. In this case, phase (C) is also asolution of all the components of the desired composition.

Instead, when phase (A) is a suspension of the hydrophobic active agentin an aqueous medium, phase (C) is also a suspension in an aqueousmedium, which will contain the dissolved soluble components such as theexcipients and surfactants.

The drying operation consists of eliminating the liquid medium, solventor dispersant, from phase (C), to obtain a dry powder with the desireddimensional features. This drying is preferably obtained by spraydrying. The features of the nozzle and the process parameters areselected so that the liquid medium is evaporated from the solution orsuspension (C) and a powder with the desired particle size is formed.

Step c) of the process for preparing the pharmaceutical compositioninstead consists of physical mixing of the powders obtained by spraydraying using the most common mixing techniques, i.e. rotating mixerssuch as Turbula, V-mixer, cylinder, double cone, cube mixers orstationary mixers used only for mixing, such as planetary, nautamix,sigma, ribbon mixers or mixer-granulators, such as Diosna. Besides thesemixers, the powders could also be mixed with devices normally used tomix liquids, such as Ultra Turrax or Silverson and, ultimately, alsoinside fluid bed granulation apparatus.

As already indicated above, a further aspect of the present invention isthat of obtaining a solid composition for use as diluent of inhalationpowders, called bulking agent (BA), comprising leucine in an amountranging from 5 to 70% by weight with respect to the weight of thepowder; a sugar in an amount ranging from 20 to 90% by weight withrespect to the weight of the powder, in which said composition has afine particle fraction (FPF) greater than 60% and a percentage of thedose delivered from the mouthpiece (DF) greater than 80%. Thiscomposition can be used as diluent of inhalation powders, i.e. as powdercapable of diluting the powder containing the active agent and at thesame time improving both the aerodynamic performance of the finalcomposition, and improving the stability of the composition.

In particular, for some active agents, with very limited doses withinthe pharmaceutical composition to be administered, it is necessary toobtain an inert powder from a pharmacological point of view, capable offacilitating the operations to divide it in the predeterminedpharmaceutical form.

The production process of the bulking agent, i.e. step b) of thepreparation process, is substantially similar to the process forpreparing the composition containing the active agent (phase a)), inparticular, this process consists of the following operations:

-   -   preparing a first phase (A) in which the leucine, the sugar and        surfactants are dissolved or dispersed in an aqueous medium;    -   drying said phase (A) in controlled conditions to obtain a dry        powder with particles with a size distribution having a median        diameter of less than 10.0 μm;    -   collecting said dry powder.

EXAMPLES

The methods for preparing the powders that constitute the pharmaceuticalcomposition and for preparing the solid composition for use as diluent(hereinafter bulking agent) of the present invention will now bedescribed.

Preparation of the Individual Powders.

The powders containing the active agents and the bulking agent wereobtained by spray drying, a drying technique used to obtain powders withuniform and amorphous particles from solutions of active agents andexcipients in appropriate solvent or mixture of solvents.

For the powders described the solvents used are water and ethyl alcoholin a fixed ratio of 70/30. The concentration of dissolved solids is 1%w/v for powders containing the active agent and 2% w/v for the bulkingagent.

In the case of the powder containing formoterol fumarate as active agentand bulking agent, all the components of the powder were dissolved inwater and the solution thus obtained was added to the portion of ethylalcohol slowly at 25° C.

For the powder containing Budesonide as active agent, the active agentwas dissolved separately in the alcohol portion to which the aqueoussolution of the excipients was added to obtain a single water-alcoholsolution.

The water-alcohol solution thus obtained was processed by means of aBuchi Mod. B290 spray dryer, using an open cycle with the followingparameters:

-   -   nozzle diameter 0.7 mm    -   atomization gas nitrogen    -   atomization pressure 4 bar    -   drying gas air    -   aspiration 100% (35 m3/h)    -   inlet temperature 170° C.    -   feed speed 8% (2.4 ml/min)    -   Powder collection system: cyclone separator with glass        collection vessel (External diameter: 8.5 cm. Height: 30.5 cm)    -   Outlet filter: nylon sleeve

At the end of the drying process the powder collection step wasperformed in controlled temperature and humidity conditions: temperature<25° C., relative humidity <35%.

The powders were packaged immediately after production in borosilicateglass vials inserted in a double aluminum foil bag heat-sealed underpartial vacuum (30%).

Preparation of the Mixtures.

The formulations described in the examples were produced by mixingpowders containing the active agents and bulking agent. Regardless ofthe quantitative ratios between initial powders, a layer-wise mixingtechnique was used, depositing the powder containing the active agentbetween two layers of bulking agent in the mixing container. The powderswere mixed using an Ultra Turrax T10 mixer for a mixing time of 5minutes considered sufficient for the 3.5 g of powder of the batchesproduced. Uniformity of the content was controlled with titre analysison 10 samples taken from different points of the bulk.

The powders were divided in sealed vials and stored inside a doublealuminum foil bag heat sealed with partial vacuum (30%).

The operations of mixing and dividing in vials were carried out inside aglove box in controlled humidity and temperature conditions; maxtemperature 20° C. and environmental relative humidity <35%.

Storage Conditions for Accelerated Stability Study.

The powders studied, packaged as described above, during the acceleratedstability study were stored in an oven at a temperature of 40° C. andrelative humidity 13%.

At each time interval established by the study, the samplescorresponding to the stability point were taken, left to cool untilreaching room temperature, opened in controlled conditions in a glovebox (temperature <20° C., RH<35%) and analyzed as established in theprotocol.

Characterization of the Powder: Particle Size Analysis.

The powders obtained were characterized in terms of dry particle sizeusing a Sympatec Helos light scattering device that analyzes theparticle size according to the Fraunhofer theory and equipped with RODOSdisperser.

The instrument was suitably calibrated with reference material andprepared following the instructions provided in the instrument usermanual.

After appropriate cleaning before analysis, an amount of powder for eachbatch produced was analyzed without any preliminary preparation of thesample.

The dispersion gas used was compressed air suitably cleansed ofparticles.

The test method specified therefore provides for compliance with thefollowing measures in relation to the sample, to the powder disperserand to the light scattering analyzer.

Sample

-   -   size: about 100 mg    -   feed procedure: with a spatula    -   pre-treatment of the sample: none

RODOS Disperser

-   -   Model M ID-NR 230 V/Hz 24 Va    -   Dispersion pressure: 3 bar

Light Scattering Analyzer

-   -   Model: Helos    -   Test method: Fraunhofer    -   Software version: Windox 4.0    -   Test lens: R1 (0.1-35 μm)    -   Minimum optical concentration: 1%    -   Activation threshold: minimum optical concentration detectable        1% for max 30 seconds of time and with at least 100 ms of        exposure of the sample.

All the tests were conducted in controlled temperature and humidityenvironments, temperature <25° C. and relative humidity <50% RH.

Size analysis provides volume median diameter (VMD) values of thepopulation of particles in the sample of powder.

Characterization of the Powder: Residual Moisture Content.

The residual moisture content in the powder was measured using the KarlFischer coulometric system method.

The C20 Compact Karl Fischer Coulometer Mettler Toledo titrator was usedfor this purpose, which uses as reagent HYDRANAL®-Coulomat AG.

The sample powders were accurately weighed in an amount of around 15-20mg and the weight was recorded in the parameters of the sample.Titration was started immediately after adding the sample to the reagentbath.

At the end of the test, the instrument indicates directly the percentageof water contained in the sample.

Characterization of the Powder: Determination of Titre and Related.

The HPLC (High Performance Liquid Chromatography) test method was usedto determine the content of the active agents and their relatedsubstances.

The test method is characterized by the following parameters:

Solvent: 50/50 methanol/phosphate buffer pH 2.7 25 mMMobile phase: acetonitrile/phosphate buffer pH 2.9 2.82 mM

-   -   gradient elution

Time % buffer Flow (min) % ACN pH 2.9 (ml/min) 0 22 78 0.5 2.5 22 78 0.53.0 41 59 0.7 8.0 41 59 0.7 10.0 70 30 0.7 12.0 22 78 0.6 15.0 22 78 0.6Injection volume: 20 μLAnalysis column: Agilent Poroshell 120 EC-C18, 100 mm×3.0 mm, 2.7 μmColumn temperature: 30° C.

Wavelength: 220 nm (Formoterol Fumarate) and 240 nm (Budesonide)

Retention time: 2.4 min (Formoterol Fumarate) and 8.0 min (Budesonide)

An HPLC Agilent model 1200 with diode array type detector, model G1315Cwas used for the test.

The samples for analysis were obtained by dissolving in the solvent anamount of powder such as to obtain a concentration of 160 μg/ml for theBudesonide and 4.5 μg/ml for the Formoterol Fumarate, as for thereference solution.

The reference solution was injected three consecutive times before thesample to determine the precision of the system expressed as relativestandard deviation percentage (RSD %), which must be less than 2%.

The active agent content is obtained by calculating the ratio of theareas with respect to the reference solution at known concentration. Thedegradation of the product is calculated as ratio between the sum of theareas of all the analysis peaks corresponding to the degradationproducts and the active agent taken as reference. All the analysis peakswhose chromatogram area was greater than 0.1% of the area of the activeagent are counted in the sum of the degradation products.

Characterization of the Powder: Differential Scanning Calorimetry.

Differential scanning calorimetry or DSC is a thermoanalytical techniqueused to determine chemical and physical phenomena with endothermic orexothermic effect in a sample, such as variations in phase, loss ofwater, chemical reactions.

In DSC the sample is heated with constant heating speed and the amountof heat required to raise its temperature is a function of its thermalcapacity. Each endothermic or exothermic phenomenon causes a reversibleor irreversible change in the thermal capacity of the material and canbe detected as a variation of the baseline of the thermogram.

Powders containing amorphous lactose show during heating a typicaldecrease in thermal capacity corresponding to the glass transition ofthe lactose from amorphous solid state to a metastable state thatrapidly leads to its crystallization, characterized by an exothermicpeak. The temperature corresponding to these phenomena varies as afunction of the composition of the sample and of the environmentalconditions in which the sample is stored and prepared.

The samples were prepared in a controlled environment (temperature <20°C., relative humidity 35-30%). 40 uL aluminum standard crucibles for DSCwere filled with a weighed amount of powder between 1 mg and 3 mg andsealed with specific lid.

Calorimetry testing of the samples in question was carried out bysubjecting the samples to a heating ramp from 20 to 200° C. with atemperature increase of 10° C./min.

The test gives a thermogram in which the thermal events that accompanyprogressive heating of the sample are visible.

The glass transition (Tg) is identifiable with a decreasing step, attimes followed by an increase in the baseline caused by relaxationenthalpy. During evaluation of the thermograms the onset temperature ofthe phenomenon (Tg onset) is calculated, regardless of the sample size.The glass transition temperature is a stability index of the powder asit is a prelude to crystallization, which takes place above 100° C. Theexothermic crystallization peak can be integrated and the area subtendedby the curve is an index of the amorphous fraction of the sample.

Characterization of the Powder: Respirability Test with MSLI.

The Multi Stage Liquid Impinger (MSLI) is a device that simulates invitro pulmonary deposition of an inhalation formulation. A inhalationformulation, delivered by appropriate inhaler and conveyed into thedevice by aspiration, is deposited in the various stages connected inseries of the impactor as a function of its aerodynamic features, suchas particle size, density, shape. Each stage of the MSLI corresponds toan interval of aerodynamic particle sizes of the powder depositedtherein and the aerodynamic size distribution of the powder is obtainedusing HPLC testing to determine the amount of active agent in eachstage, making it possible to calculate the mass median aerodynamicdiameter (MMAD) and the respirable fraction (also known as fine particlefraction, FPF), considered according to the European Pharmacopoeia withaerodynamic diameter <5.0 μm.

For the respirability test, the powders of the formulations of theexamples were partitioned into Size 3 HPMC capsules and delivered fromRS01 powder inhaler—model 7 monodose, code 239700001AB(Aerolizer-Plastiape S.p.A.).

The device was assembled following the instructions for use and theindications of the European Pharmacopoeia.

For test purposes, it is necessary to deliver 10 powder capsules foreach respirability test. The tests were conducted at a flow of 60 Lpmfor 4 seconds deriving from a pressure drop of 2 KPa in the system.

The following aerodynamic diameter cut-offs correspond to this flowvalue for each stage.

-   -   stage 1: >13 μm    -   stage 2: from 13 μm to 6.8 μm    -   stage 3: from 6.8 μm to 3.1 μm    -   stage 4: from 3.1 μm to 1.7 μm    -   stage 5 (filter): <1.7 μm

The respirable fraction (Fine Particle Fraction) comprises particleswith aerodynamic diameter of less than 5 μm and is calculated usingspecific software (CITDAS Copley).

The aerodynamic parameters of an inhalation formulation subjected toMSLI analysis are expressed in terms of:

-   -   Delivered Fraction (DF): i.e. the percentage of the dose of        active agent delivered from the mouthpiece of the inhaler    -   Fine Particle Dose (FPD): respirable dose of active agent,        having aerodynamic diameter <5.0 μm.    -   Fine Particle Fraction (FPF): respirable fraction (aerodynamic        diameter <5.0 μm) of active agent expressed as percentage of the        amount delivered.    -   Mass Median Aerodynamic Diameter (MMAD): median aerodynamic        diameter of the particles delivered.

Quantitative determination of the active agent in each stage wasperformed by HPLC using the test method for content and degradationproducts.

Example 1

Example 1 was conducted producing powders containing FormoterolFumarate, which is an active agent sensitive to the presence of freewater in the formulation.

Together with formoterol powders, powders containing different amountsof leucine and lactose or mannitol were produced.

The example highlights the protective effect of lactose againstformoterol, this protective effect is explained considering that lactoseis capable of exerting a scavenger effect against free water present inthe formulation.

To demonstrate this, powders of 3 types were produced:

-   -   A powder containing exclusively formoterol and leucine    -   2 powders with different lactose contents together with        formoterol and leucine    -   2 powders in which lactose was substituted by a different        pharmaceutical excipient widely utilized by spray drying:        mannitol

The powders with lactose tend to acquire moisture over time, withconsequent decrease of Tg, but degradation over time is limited. Thislimited degradation is presumably due to a scavenger effect produced bythe lactose against the water, which is thus trapped in a rigidstructure and prevented from reacting with the other components.Differently, the powders without lactose which was already crystalline,undergoes chemical degradation.

Of the two powders containing lactose, the one with 50% is better, as itis more stable over time.

TABLE 1A Water Formoterol content (%) Ex. Active (%) Leucine % Sugar T0T28 days 1 Formoterol 5 95 NO Sugar 0.9 0.9 2 Formoterol 5 70 Lactose1.4 1.8 3 Formoterol 5 45 Lactose 2.1 2.7 4 Formoterol 5 70 Mannitol 0.90.9 5 Formoterol 5 45 Mannitol 1.0 0.9

TABLE 1B P. size Degradation Tg (° C.) (VMD) products (%) Ex. T0 T28days T0 T28 days T0 T28 days 1 Not detected Not detected 2.6 2.7 0.6 0.92 62.7 56.9 2.0 1.9 0.4 0.4 3 66.3 57.5 1.6 1.6 0.3 0.3 4 Not detectedNot detected 2.3 2.2 0.2 1.6 5 Not detected Not detected 1.6 1.6 0.1 1.4

Example 2

The example was conducted producing powders containing Budesonide asactive agent (defined as HLSA Bud in the table) formulated with lactoseand leucine at two different quantitative levels.

Following preparation of the powders of example 6 and 9, these weremixed with 2 types of bulking agent powders (defined as BA in thetable), i.e. of powder containing leucine and lactose but with no activeagent.

The presence of leucine in different amounts ranging from 0% to 20%serves to highlight the properties of disaggregating agent that thistakes place in the formulation, with positive effects on parameters suchas Delivered Fraction and Fine Particle Fraction.

This second part of the study highlights the ability of the BulkingAgent to promote complete emptying of the capsule.

Nevertheless, the composition of the Bulking agent is critical, since aBulking Agent that has too much leucine produces effects of chemicaldegradation of the active agent.

According to the present invention, the powder is acceptable, i.e. isconsidered within the optimal parameters for inhalation administration,when:

-   -   the degradation products are less than 0.5% on the total of        active agent, at the time T2 (degradation products T2<0.5% tot);    -   the Delivered Fraction, i.e. the percentage of the dose of        active agent delivered from the mouthpiece of the inhaler, is        greater than 90% at the time T2 (ED % T2>90%);    -   the Fine Particle Fraction, i.e. the amount of fine particles        below 5 μm, is greater than 60% at the time T2. (FPT T2>60%).

TABLE 2A Powder HLSA Bud (composition %) BA (composition %) mixtureTween Tween HLSA Bud BA Ex. Active Bud. Leucine Lactose 80 LeucineLactose 80 (mg) (mg) 6 Budesonide 8 0.0 91.5 0.5 5.0 0.0 7 Budesonide 80.0 91.5 0.5 50.0 49.5 0.5 5.0 15.0 8 Budesonide 8 0.0 91.5 0.5 99.5 0.00.5 5.0 15.0 9 Budesonide 8 20.0 71.5 0.5 5.0 0.0 10 Budesonide 8 20.071.5 0.5 50.0 49.5 0.5 5.0 15.0 11 Budesonide 8 20.0 71.5 0.5 99.5 0.00.5 5.0 15.0

TABLE 2B Water content (%) Particle Size (μm) Active content % Ex. T0 T1T2 T0 T1 T2 T0 T1 T2 6 2.8 3.2 2.9 1.9 2.1 2.6 101.2 96.7 102.5 7 2.63.2 3.1 1.8 1.9 2.2 105.7 104.1 105.6 8 0.8 1.6 1.1 2.6 2.7 2.8 101101.3 101.1 9 2.5 3.6 3 1.5 1.5 1.7 99.2 100.9 100.6 10 2.6 3.2 2.8 1.71.7 1.8 100.5 100.7 100.7 11 0.7 1.2 1.3 2.7 2.7 2.8 99.3 101.5 100.7

The active content measured after 1 and 2 months was variable but alwaysbetween 95 and 110% of the theoretical content.

TABLE 2C Degradation (%) DF (%) FPF (%) Ex. T0 T1 T2 T0 T1 T2 T0 T1 T2 60.0 0.2 0.2 81.8 81.4 84.6 51.2 38.0 36.7 7 0.1 0.2 0.3 95.4 94.6 97.144.6 40.0 42.6 8 0.0 1.2 1.9 94.9 95.4 94.9 34.4 46.7 43.4 9 0.2 0.2 0.385.2 84.1 85.2 71.0 78.4 73.6 10 0.2 0.3 0.3 93.8 95.8 95.2 70.5 71.465.8 11 0.2 1.1 1.8 93.4 96.4 93.9 42.1 62.0 63.3

Example 3

Example 3 was conducted expanding on Example 2 varying the amounts ofleucine and sugar in the powders containing Budesonide as active agent(defined as HLSA Bud in the table). Together with the powders containingBudesonide, bulking agent powders were produced containing lactose andleucine at three different levels of leucine using lactose as filler toform the bulking agent (defined as BA in the table) i.e. of powdercontaining leucine and lactose but with no active agent.

The presence of leucine in three different amounts 0%, 50% and 91.5%serves to highlight the properties of disaggregating agent that thistakes in the formulation with positive effects on parameters such asDelivered Fraction and Fine Particle Fraction.

Following preparation of the powders of Examples 12, 13 and 14, thesewere mixed with 3 types of Bulking Agent powders.

These 3 Bulking Agents also contain Leucine in three different amounts(0%, 50% and 99.5%)

This further part of the study highlights the capacity of the Bulkingagent to promote complete emptying of the capsule.

Nevertheless, the composition of the Bulking agent is critical since aBulking Agent with too much leucine produces effects of chemicaldegradation of the active agent.

According to the present invention the powder is acceptable, i.e. isconsidered within the optimal parameters for inhalation administration,when:

-   -   the degradation products are less than 1% of the total active        agent, at the time T0 (degradation products T0<1% tot);    -   the Delivered Fraction (DF), i.e. the percentage of the dose of        active agent delivered from the mouthpiece of the inhaler, is        greater than 80% at the time T3 (ED % T3>80%);    -   the Fine Particle Fraction, i.e. the amount of fine particles        with aerodynamic diameter of less than 5.0 μm, is greater than        60% at the time T0 and at the time T3 (FPF T0 and T3>60%).

TABLE 3A HLSA Bud (composition %) BA (composition %) Powder mixtureTween Tween HLSA Bud BA Ex. Active Bud. Leucine Lactose 80 LeucineLactose 80 (mg) (mg) 12 Budesonide 8 0.0 91.5 0.5 5.0 0 13 Budesonide 850.0 41.5 0.5 5.0 0 14 Budesonide 8 91.5 0.0 0.5 5.0 0 15 Budesonide 80.0 91.5 0.5 0.0 99.5 0.5 0.1 9.9 16 Budesonide 8 0.0 91.5 0.5 50.0 49.50.5 0.1 9.9 17 Budesonide 8 0.0 91.5 0.5 99.5 0.0 0.5 0.1 9.9 18Budesonide 8 50.0 41.5 0.5 0.0 99.5 0.5 0.1 9.9 19 Budesonide 8 50.041.5 0.5 50.0 49.5 0.5 0.1 9.9 20 Budesonide 8 50.0 41.5 0.5 99.5 0.00.5 0.1 9.9 21 Budesonide 8 91.5 0.0 0.5 0.0 99.5 0.5 0.1 9.9 22Budesonide 8 91.5 0.0 0.5 50.0 49.5 0.5 0.1 9.9 23 Budesonide 8 91.5 0.00.5 99.5 0.0 0.5 0.1 9.9

TABLE 3B Water Particle Active content (%) Size (μm) content % Ex. T0 T3T0 T3 T0 T3 12 2.6 2.3 2.0 2.2 102.9 102.3 13 1.9 1.6 1.9 1.9 101.4 99.314 0.7 0.4 3.0 3.0 89.3 91.6 15 2.6 1.9 3.1 4.4 95.9 101.9 16 2.2 2.02.1 1.9 101.3 104.6 17 1.0 0.4 3.2 3.7 103.4 100.2 18 2.7 1.7 2.9 4.5102.2 95.3 19 2.6 2.1 2.0 2.0 99.3 103.1 20 0.9 0.5 3.2 3.4 92.9 83.4 212.9 1.9 3.6 3.8 98.8 89 22 2.3 2.3 2.4 2.4 99.8 92.6 23 0.4 0.4 3.3 3.591.4 62.8

TABLE 3C Degradation (%) DF (%) FPF (%) Ex. T0 T3 Growth T0 T3 T0 T3 120.0 0.0 0.0 73.7 73.6 45.8 37.9 13 0.4 0.7 0.3 79.1 79.0 67.6 74.4 141.6 4.4 2.8 92.6 93.1 69.6 78.5 15 0.0 0.4 0.4 94.3 94.6 35.5 24.0 160.0 0.4 0.4 92.9 94.7 44.1 40.0 17 0.0 1.9 1.9 96 96.0 44.3 33.7 18 0.40.7 0.3 95.6 95.6 44.3 27.2 19 0.4 1.5 1.1 94.4 95.5 64.6 75.2 20 0.413.2 12.8 96 95.8 57.5 65.6 21 1.7 3.0 1.3 95.9 95.6 47.2 18.5 22 1.75.6 3.9 92.3 95.7 51.3 72.0 23 1.8 23.7 21.9 95.8 97.0 47.2 79.4

Example 4

The example was conducted by producing powders containing FormoterolFumarate as active agent (defined as HLSA FF in the table) formulatedwith lactose and leucine in two different amounts.

Together with the powders containing Formoterol Fumarate, powders wereproduced containing lactose and leucine in three different amounts ofleucine using lactose as filler to form a bulking agent (defined as BAin the table), i.e. of powder containing leucine and lactose but with noactive agent.

The presence of leucine in three different amounts 0%, 50% and 91.5%serves to highlight the properties of disaggregating agent that thistakes in the formulation with positive effects on parameters such asDelivered Fraction and Fine Particle Fraction.

Following preparation of the powders of Examples 12, 13 and 14, thesewere mixed with 3 types of Bulking Agent powders.

These 3 Bulking Agents also contain Leucine in three different amounts(0%, 50% and 99.5%).

This further part of the study highlights the capacity of the Bulkingagent to promote complete emptying of the capsule.

Nevertheless, the composition of the Bulking agent is critical since aBulking Agent that has too much leucine produces effects of chemicaldegradation of the active agent.

According to the present invention the powder is acceptable, i.e. isconsidered within the optimal parameters for inhalation administration,when:

-   -   the degradation products are less than 1% of the total active        agent, at the time T0 (degradation products T0<1% tot);    -   the Delivered Fraction (DF), i.e. the percentage of the dose of        active agent delivered from the mouthpiece of the inhaler, is        greater than 80% at the time T3 (ED % T3>80%);    -   the Fine Particle Fraction, i.e. the amount of fine particles        less than 5.0 μm, is greater than 60% at the time T0 and at the        time T3 (FPF T0 and T3>60%).

TABLE 4A Powder mixture HLSA FF (composition %) BA (composition %) HLSABud BA Ex Active Formoterol Leucine Lactose Tween 80 Leucine LactoseTween 80 (mg) (mg) 24 Formoterol 2.25 0.0 97.25 0.5 5.0 0.0 25Formoterol 2.25 20.0 77.25 0.5 5.0 0.0 26 Formoterol 2.25 97.25 0.0 0.55.0 0.0 27 Formoterol 2.25 0.0 97.25 0.5 0.0 99.5 0.5 0.01 9.99 28Formoterol 2.25 0.0 97.25 0.5 50.0 49.5 0.5 0.01 9.99 29 Formoterol 2.250.0 97.25 0.5 99.5 0.0 0.5 0.01 9.99 30 Formoterol 2.25 20.0 77.25 0.50.0 99.5 0.5 0.01 9.99 31 Formoterol 2.25 20.0 77.25 0.5 50.0 49.5 0.50.01 9.99 32 Formoterol 2.25 20.0 77.25 0.5 99.5 0.0 0.5 0.01 9.99 33Formoterol 2.25 97.25 0.0 0.5 0.0 99.5 0.5 0.01 9.99 34 Formoterol 2.2597.25 0.0 0.5 50.0 49.5 0.5 0.01 9.99 35 Formoterol 2.25 97.25 0.0 0.599.5 0.0 0.5 0.01 9.99

TABLE 4B Water Particle Active content (%) Size (μm) content % Ex. T0 T3T0 T3 T0 T3 24 4.2 3.6 2.5 2.85 96.6 97.4 25 3.3 3.3 1.5 1.33 100.3 95.326 0.8 0.6 2.6 2.59 95.2 89.3 27 2.8 1.7 3.4 3.98 98.8 90.5 28 3.2 2 2.02.12 98.5 97 29 0.7 0.3 3.3 3.59 95.5 86.1 30 2.6 1.8 3.1 3.88 97.2 88.931 2.4 1.7 2.1 2.16 96.8 101.5 32 0.6 0.4 2.8 3.52 92.7 76.5 33 2.6 2.33.3 3.82 78.7 73.2 34 2.4 1.8 2.1 2.17 84.6 87.8 35 0.4 0.2 3.2 3.5293.6 68.8

TABLE 4C Degradation (%) DF (%) FPF (%) Ex. T0 T3 Growth T0 T3 T0 T3 240.8 0.7 0.0 76.8 79.2 38.9 42.7 25 0.2 0.9 0.7 78.3 79.1 71.9 70.6 261.0 6.9 5.9 94.1 95.7 77.8 87.3 27 0.8 0.5 0.0 93.5 90.7 36.9 32.9 281.0 0.7 0.0 85.7 81.3 37.5 48.2 29 1.6 6.6 5 96.8 93.9 30.6 37.8 30 0.23.9 3.7 96.1 91.8 38 29.8 31 0.2 0.6 0.4 91.4 92.2 73.4 78.1 32 1.3 7.46.1 96.6 94 65.1 69.3 33 0.7 5.5 4.8 95 93 39.3 30.8 34 0.8 2.4 1.6 90.197.7 45.3 78.9 35 2.3 12.8 10.5 95.5 97.2 71.1 68.3

Example 5

The example was conducted producing formulations containing tiotropiumbromide powders alone (defined as HLSA.Tio and standing for high loadingsingle active of tiotropium) or powders containing a combination oftiotropium bromide together with formoterol fumarate as active agents(defined as HLDA.TioFF and standing for high loading double active oftiotropium and formoterol).

Together with the powders containing tiotropium or tiotropium andformoterol, a bulking agent powder was produced and obtainedincorporating leucine, lactose and tween 80.

Following the preparation of the HLSA.Tio and the HLDA.TioFF these weremixed at two different dosing levels with different amounts of Bulkingagent.

According to the present invention the powders are acceptable and withinthe acceptable parameters for optimal inhalation administration:

-   -   the Delivered Fraction (DF), i.e. the percentage of the dose of        active agent delivered from the mouthpiece of the inhaler, is        greater than 80% at the time T0 0;    -   the Fine Particle Fraction, i.e. the amount of fine particles        less than 5.0 μm, is greater than 60% at the time T0.

TABLE 5A Powder mixture HLSA.Tio/HLDA.TioFF (composition %) BA(composition %) HLSA/HLDA Ex Active Active Tiotropium Formoterol LeucineLactose Tween 80 Leucine Lactose Tween 80 (mg) BA (mg) 36 Titropium 3.020.0 76.5 0.5 50.0 49.5 0.5 0.6 2.4 37 Titropium Formoterol 3.0 3.0 20.073.5 0.5 50.0 49.5 0.5 0.6 2.4 38 Titropium 3.0 20.0 76.5 0.5 50.0 49.50.5 0.1 9.9 39 Titropium Formoterol 3.0 3.0 20.0 73.5 0.5 50.0 49.5 0.50.1 9.9

TABLE 5B Water content Particle (%) size (μm) EX T0 T0 36 2.7 1.7 37 2.31.4 38 1.8 1.8 39 2.6 1.5

TABLE 5C DF (%) DF (%) FPF (%) FPF (%) Tiotropium Formoterol TiotropiumFormoterol EX T0 T0 T0 T0 36 85.1 62.4 37 87.4 88.4 61.3 64.3 38 92.468.7 39 93.2 93.1 69.5 66.7

1. A pharmaceutical composition for inhalatory use in powder form,characterized by comprising at least a first and a second powder,wherein at least said first powder comprises an active principle in anamount greater than 1% by weight of said first powder, said first andsecond powder comprising: a) leucine in amount from 5 to 70% by weightof each powder; b) a sugar in amount from 20 to 90% by weight of eachpowder; wherein said composition has a fine particle fraction (FPF)greater than 60% and an delivered fraction (DF) greater than 80%. 2.Composition according to claim 1, wherein the ratio from the amount ofpowder in amorphous form which form the composition expressed by weight,to the amount of sugar in the composition expressed in weight, is from0.8 to 1.5.
 3. Composition according to claim 1, wherein said first andsecond powder comprise a surfactant in an amount from 0.2 to 2% byweight of each powder.
 4. Composition according to claim 1, wherein saidactive principle is an hydrolysable active principle.
 5. Compositionaccording to claim 1, wherein said active principle is selected from thegroup consisting of: inhalation bronchodilators with short and longduration of action, corticosteroids, anticholinergics, antibiotics,mucolytics, heparin and its derivatives, and antioxidant substances. 6.Composition according to claim 1, wherein said antioxidant substancesare selected from the group consisting of: N-acetylcysteine, Carnosine,Melatonin, Resveratrol, Ascorbic Acid, Alpha-tocopherol, Folic Acid,Trans-Caffeic Acid, Hesperidin, Epigallocatechin Gallate, Delphinidin,Acid Rosmainico, Myricetin, 5-methyltetrahydrofolic Acid,5-formyltetrahydrofolic Acid, Astaxanthin, Lycopene, and Curcumin. 7.Composition according to claim 1, wherein said sugar is selected fromthe group consisting of: lactose, trehalose, sucrose and maltodextrin.8. Composition according to claim 1, wherein said leucine is in aquantity from 15 to 70% by weight of each powder.
 9. Compositionaccording to claim 1, wherein said leucine is in a quantity from 18 to55% by weight of each powder.
 10. Composition according to claim 1,wherein said sugar is in an amount from 20 to 80% by weight of eachpowder.
 11. Composition according to claim 1, wherein said sugar is inan amount from 40 to 80% by weight of each powder.
 12. Compositionaccording to claim 1, wherein said surfactant is selected from the groupconsisting of: benzalkonium chloride, cetrimide, docusate sodium,glyceryl monooleate, sorbitan esters, sodium lauryl sulfate,polysorbates, phospholipids, bile salts, polysorbates, block copolymersof polyoxyethylene and polyoxypropylene.
 13. Composition according toclaim 1, wherein said surfactant is in an amount from 0.4 to 0.8% byweight of each powder.
 14. Composition according to claim 1, whereincomprising a third powder comprising an active ingredient in an amountgreater than 1% by weight of said third powder, leucine in amount from 5to 70% by weight of said third powder, a sugar in an amount from 20 to90% by weight of said third powder.
 15. Composition according to claim1, wherein said powders have a X50 less than 5 μm.
 16. Process forpreparing a pharmaceutical composition according to claim 1, whichcomprises the following steps: a) providing at least a first powderobtained by spray draying comprising an active principle in an amountgreater than 1% by weight of the powder, leucine in a quantity from 5 to70% by weight of the powder, a sugar substantially amorphous after theobtaining of the powder with spray-drying in a quantity from 20 to 90%by weight of the poor; b) providing a second powder obtained by spraydraying comprising leucine in an amount comprised from 5 to 70% byweight of the powder, a sugar substantially amorphous after theobtaining of the powder with spray-drying in a quantity comprised from20 to 90% by weight of the powder; c) mixing the powders.
 17. Solidcomposition for use as a diluent of inhalatory powders comprising apowder, it which comprises; a) leucine in amount from 5 to 70% by weightof each powder; b) a sugar in amount from 20 to 90% by weight of eachpowder; wherein said composition has a fine particle fraction (FPF)greater than 60% and an emitted dose (ED) greater than 80%.
 18. Kit forthe administration of a drug as inhalatory powder, comprising a meteredamount of the composition according to claim 1 and a device forinhalation.
 19. Composition according to claim 2, wherein said first andsecond powder comprise a surfactant in an amount from 0.2 to 2% byweight of each powder.
 20. Composition according to claim 2, whereinsaid active principle is an hydrolysable active principle.